Fine‐tuning of defences and counter‐defences in a specialised plant–herbivore system

1. The plant–herbivore arms race has been postulated to be a major driver for generating biological and biochemical diversity on Earth. Herbivore feeding is reduced by the production of chemical and physical barriers, but increases plant resistance against subsequent attack. Accordingly, specialisation is predicted to be an outcome of herbivores being able to circumvent plant‐induced defences. 2. Using a specialised plant–herbivore system, in which adult chrysomelid beetles (Chrysochus auratus) feed on leaves and larvae feed on roots of dogbane (Apocynum spp.), this study investigated whether root latex and cardenolides are effective against the soil‐dwelling larvae, and whether such defences could be circumvented by the herbivore. 3. Across two Apocynum species, C. auratus larvae were not affected by latex production or cardenolide amounts and diversity. By contrast, cardenolide apolarity was detrimental to larval growth. Yet larval feeding decreased average root cardenolide apolarity in A. cannabinum and larvae performed better on those plants. Finally, above‐ground induction rendered the plants more toxic by increasing root cardenolide apolarity and maintaining it, even during subsequent larval herbivory. 4. Therefore, the intimate relationship and interaction between Chrysochus and Apocynum are maintained by a delicate balance of herbivore manipulation and plant chemical induction.     

Effects of plant nutrition on the balance of insect relevant cardenolides and glucosinolates in Erysimum cheiranthoides

The possible effects of environmental stress on plant chemistry that are important to herbivorous insects were examined by growing a wild crucifer, Erysimum cheiranthoides, under different nutrient regimes. Oviposition by the cabbage butterfly, Pieris rapae, is thought to be affected by the balance of glucosinolates (stimulants) and cardenolides (deterrents) at the surface of leaves. E. cheiranthoides seedlings were provided with three levels of nitrogen and two levels of sulfur for a period of 15 days before analysis of semiochemicals in whole leaf tissue and at the surface of the foliage. The ratio of cardenolides to glucosinolates in the plants at elevated C/N ratios followed the carbon/nutrient balance hypothesis. However, a high nitrogen supply enhanced biomass production to the extent that concentrations of secondary compounds were unchanged or reduced. The concentration of glucosinolates (glucoiberin and glucocheirolin) at the surface was positively related to whole tissue levels. However, cardenolide (erysimoside and erychroside) concentrations, which were highest in leaf tissue of nitrogen-deficient plants, had the lowest surface levels on foliage of these plants. Possible reasons for differential expression of cardenolides and glucosinolates in a plant as a result of nutrient deficiency are discussed.     

Fitness costs of chemical defense in Plantago lanceolata L.: effects of nutrient and competition stress

Fitness costs of defense are often invoked to explain the maintenance of genetic variation in levels of chemical defense compounds in natural plant populations. We investigated fitness costs of iridoid glycosides (IGs), terpenoid compounds that strongly deter generalist insect herbivores, in ribwort plantain (Plantago lanceolata L.) using lines that had been artificially selected for high and low leaf IG concentrations for four generations. Twelve maternal half-sib families from each selection line were grown in four environments, consisting of two nutrient and two competition treatments. We tested whether: (1) in the absence of herbivores and pathogens, plants from lines selected for high IG levels have a lower fitness than plants selected for low IG levels; and (2) costs of chemical defense increase with environmental stress. Vegetative biomass did not differ between selection lines, but plants selected for high IG levels produced fewer inflorescences and had a significantly lower reproductive dry weight than plants selected for low IG levels, indicating a fitness cost of IG production. Line-by-nutrient and line-by-competition interactions were not significant for any of the fitness-related traits. Hence, there was no evidence that fitness costs increased with environmental stress. Two factors may have contributed to the absence of higher costs under environmental stress. First, IGs are carbon-based chemicals. Under nutrient limitation, the relative carbon excess may result in the production of IGs without imposing a further constraint on growth and reproduction. Second, correlated responses to selection on IG levels indicate the existence of a positive genetic association between IG level and cotyledon size. At low nutrient level, a path analysis based on family means revealed that in the presence of competitors, the negative direct effect of a high IG level on aboveground plant dry weight was partly offset by a positive direct effect of the associated larger cotyledon size. This indicates that fitness costs of defense may be modulated by environment-specific fitness effects of genetically associated traits.     

Molecular adaptation of Chrysochus leaf beetles to toxic compounds in their food plants

Herbivores that feed on toxic plants must overcome plant defenses and occasionally may even benefit from them. The current challenge is to understand how herbivores evolve the necessary physiological adaptations and which changes at the molecular level are involved. In this context we studied the leaf beetles genus Chrysochus (Coleoptera, Chrysomelidae). Two species of this genus, C. auratus and C. cobaltinus, feed on plants that contain toxic cardenolides. These beetles not only avoid poisoning by the toxin but also use it for their own defense against predators. All other Chrysochus species feed on plants that are devoid of cardenolides. The most important active principle of cardenolides is their capacity to bind to and thereby block the ubiquitous Na(+)/K(+)-ATPase responsible for maintaining cellular potentials. By analyzing the DNA sequence of the putative ouabain-binding site of the alpha-subunit of the Na(+)/K(+)-ATPase gene of Chrysochus and its close relatives feeding on plants with or without cardenolides, we here trace the evolution of cardenolide insensitivity in this group of beetles. The most interesting difference among the sequences involves the amino acid at position 122. Whereas all species that do not encounter cardenolides have an asparagine in this position, both Chrysochus species that feed on cardenolide plants have a histidine instead. This single amino acid substitution has already been shown to confer cardenolide insensitivity in the monarch butterfly. A mtDNA-based phylogeny corroborates the hypothesis that the asparagine at position 122 of the alpha-subunit of the Na(+)/K(+)-ATPase gene as observed in Drosophila and other insects is the plesiomorphic condition in this group of leaf beetles. The later host-plant switch to cardenolide-containing plants in the common ancestor of C. auratus and C. cobaltinus coincides with the exchange of the asparagine for a histidine in the ouabain binding site.     

Effects of soil nutrients on the sequestration of plant defence chemicals by the specialist insect herbivore,Danaus plexippus

1. Although anthropogenic nitrogen (N) enrichment has significantly changed the growth, survival and reproduction of herbivorous insects, its effects on the defensive sequestration of secondary chemicals by insect herbivores are less well understood. Previous studies have shown that soil nutrient availability can affect sequestration directly through changing concentrations of plant defence chemicals, or indirectly through altering growth rates of herbivores. There has been less exploration of how nutrient deposition affects the consumption of secondary chemicals and subsequent sequestration efficiency. In the current study, the overall effect of soil N availability on cardenolide sequestration by the monarch caterpillar Danaus plexippus was examined. Specifically, the effects of soil nutrient availability on growth, consumption, excretion and sequestration efficiency of cardenolides by D. plexippus larvae fed on the tropical milkweed Asclepias curassavica were measured. 2. The results showed that soil N and phosphorus (P) fertilisation significantly reduced caterpillar growth rate and the sequestration efficiency of cardenolides by monarch caterpillars feeding on A. curassavica. The lowered sequestration efficiency was accompanied by higher concentrations of cardenolides in frass. Although the total cardenolide contents of caterpillars were lower under high N or P fertilisation levels, caterpillar cardenolide concentrations were constant across fertilisation treatments because of lower growth rates (and therefore lower body mass) under high fertilisation. It is concluded that anthropogenic N deposition may have multiple effects on insect herbivores, including their ability to defend themselves from predators with sequestered plant defences.     

Evolution of latex and its constituent defensive chemistry in milkweeds (Asclepias): a phylogenetic test of plant defense escalation

A tremendous diversity of plants exude sticky and toxic latex upon tissue damage, and its production has been widely studied as a defensive adaptation against insect herbivores. Here, we address variation in latex production and its constituent chemical properties (cardenolides and cysteine proteases) in 53 milkweeds [Asclepias spp. (Apocynaceae)], employing a phylogenetic approach to test macroevolutionary hypotheses of defense evolution. Species were highly variable for all three traits, and they showed little evidence for strong phylogenetic conservatism. Latex production and the constituent chemical defenses are thus evolutionarily labile and may evolve rapidly. Nonetheless, in phylogenetically independent analyses, we show that the three traits show some correlations (and thus share a correlated evolutionary history), including a positive correlation between latex exudation and cysteine protease activity. Conversely, latex exudation and cysteine protease activity both showed a trade‐off with cardenolide concentrations in latex. We also tested whether these traits have increased in their phenotypic values as the milkweeds diversified, as predicted by plant defense escalation theory. Alternative methods of testing this prediction gave conflicting results – there was an overall negative correlation between amount of evolutionary change and amount of latex exudation; however, ancestral state reconstructions indicated that most speciation events were associated with increases in latex. We conclude by (i) summarizing the evidence of milkweed latex itself as a multivariate defense including the amount exuded and toxin concentrations within, (ii) assessing the coordinated evolution of latex traits and how this fits with our previous notion of ‘plant defense syndromes’, and finally, (iii) proposing a novel hypothesis that includes an ‘evolving community of herbivores’ that may promote the escalation or decline of particular defensive strategies as plant lineages diversify.     

Effects of cross host species inoculation of nitrogen‐fixing endophytes on growth and leaf physiology of maize

Plants that grow and thrive under abiotic stress often do so with the help of endophytic microorganisms. Although nitrogen‐fixing (diazotrophic) endophytes colonize many wild plants, these natural relationships may be disrupted in cultivated crop species where breeding and genotype selection often occur under conditions of intensive fertilization and irrigation. Many energy crops including corn may still benefit from diazotrophic endophyte inoculations allowing for more efficient biomass production with less input of petroleum‐derived fertilizer. A selection of diazotrophic endophytes isolated from willow (Salix sitchensis, Sitka willow) and poplar (Populus trichocarpa, black cottonwood) growing in nutrient‐poor river sides were used as inoculum in three experiments testing the effect on plant growth and leaf level physiology of a sweet corn variety under various levels of applied nitrogen fertilizer. We report substantial growth promotion with improved leaf physiology of corn plants in response to diazotrophic endophyte inoculations. Significant gains of early biomass with a greater root : shoot ratio were found for plants receiving endophytic inocula over the uninoculated control groups regardless of the nitrogen level. Furthermore, inoculated plants exhibited consistently higher rates of net CO₂ assimilation than did those without endophytic inoculation. These results have beneficial implications for enhanced plant growth in a low‐input system on nutrient‐poor sites. The immediate increase of root mass observed in endophyte inoculated plants has the potential to provide better establishment and early growth in resource‐limited environments. The initial results of this study also indicate that the beneficial effect from endophytes isolated from poplar and willow species is not restricted to the species from which they were initially isolated.     

The demographic effects of functional traits: an integral projection model approach reveals population‐level consequences of reproduction‐defence trade‐offs

Quantitatively linking individual variation in functional traits to demography is a necessary step to advance our understanding of trait‐based ecological processes. We constructed a population model for Asclepias syriaca to identify how functional traits affect vital rates and population growth and whether trade‐offs in chemical defence and demography alter population growth. Plants with higher foliar cardenolides had lower fibre, cellulose and lignin levels, as well as decreased sexual and clonal reproduction. Average cardenolide concentrations had the strongest effect on population growth. In both the sexual and clonal pathway, the trade‐off between reproduction and defence affected population growth. We found that both increasing the mean of the distribution of individual plant values for cardenolides and herbivory decreased population growth. However, increasing the variance in both defence and herbivory increased population growth. Functional traits can impact population growth and quantifying individual‐level variation in traits should be included in assessments of population‐level processes.     

Gene duplications circumvent trade‐offs in enzyme function: Insect adaptation to toxic host plants

Herbivorous insects and their adaptations against plant toxins provide striking opportunities to investigate the genetic basis of traits involved in coevolutionary interactions. Target site insensitivity to cardenolides has evolved convergently across six orders of insects, involving identical substitutions in the Na,K‐ATPase gene and repeated convergent gene duplications. The large milkweed bug, Oncopeltus fasciatus, has three copies of the Na,K‐ATPase α‐subunit gene that bear differing numbers of amino acid substitutions in the binding pocket for cardenolides. To analyze the effect of these substitutions on cardenolide resistance and to infer possible trade‐offs in gene function, we expressed the cardenolide‐sensitive Na,K‐ATPase of Drosophila melanogaster in vitro and introduced four distinct combinations of substitutions observed in the three gene copies of O. fasciatus. With an increasing number of substitutions, the sensitivity of the Na,K‐ATPase to a standard cardenolide decreased in a stepwise manner. At the same time, the enzyme's overall activity decreased significantly with increasing cardenolide resistance and only the least substituted mimic of the Na,K‐ATPase α1C copy maintained activity similar to the wild‐type enzyme. Our results suggest that the Na,K‐ATPase copies in O. fasciatus have diverged in function, enabling specific adaptations to dietary cardenolides while maintaining the functionality of this critical ion carrier.     

Chemical defense,mycorrhizal colonization and growth responses in <Emphasis Type="Italic">Plantago lanceolata</Emphasis> L.

Allelochemicals defend plants against herbivore and pathogen attack aboveground and belowground. Whether such plant defenses incur ecological costs by reducing benefits from plant mutualistic symbionts is largely unknown. We explored a potential trade-off between inherent plant chemical defense and belowground mutualism with arbuscular mycorrhizal fungi (AMF) in Plantago lanceolata L., using plant genotypes from lines selected for low and high constitutive levels of the iridoid glycosides (IG) aucubin and catalpol. As selection was based on IG concentrations in leaves, we first examined whether IG concentrations covaried in roots. Root and leaf IG concentrations were strongly positively correlated among genotypes, indicating genetic interdependence of leaf and root defense. We then found that root AMF arbuscule colonization was negatively correlated with root aucubin concentration. This negative correlation was observed both in plants grown with monocultures of Glomus intraradices and in plants colonized from whole-field soil inoculum. Overall, AMF did not affect total biomass of plants; an enhancement of initial shoot biomass was offset by a lower root biomass and reduced regrowth after defoliation. Although the precise effects of AMF on plant biomass varied among genotypes, plants with high IG levels and low AMF arbuscule colonization in roots did not produce less biomass than plants with low IG and high AMF arbuscule colonization. Therefore, although an apparent trade-off was observed between high root chemical defense and AMF arbuscule colonization, this did not negatively affect the growth responses of the plants to AMF. Interestingly, AMF induced an increase in root aucubin concentration in the high root IG genotype of P. lanceolata. We conclude that AMF does not necessarily stimulate plant growth, that direct plant defense by secondary metabolites does not necessarily reduce potential benefits from AMF, and that AMF can enhance concentrations of root chemical defenses, but that these responses are plant genotype-dependent.     

Dependence of starch storage on nutrient availability and photon flux density in small birch Betula pendula Roth)

Abstract Small birch plants (Betula pendula Roth) were grown in a climate chamber at different levels of nutrient availability and at two photon flux densities. The extent to which starch storage was dependent upon nutrient availability and photon flux density was investigated. Acclimated values of starch concentration in leaves were highest at low nutrient availability and high photon flux density. Starch storage in roots was only found at the lowest nutrient availability. However, the relative rate of starch storage (starch stored per unit plant dry weight and time) was higher in plants with good nutrition. The data suggest that, at sub-optimal nutrient availability, the momentary rate of net shoot photosynthesis is unlikely to limit the structural (as opposed to carbon storage) growth of the plant. Although photosynthetic rate per unit leaf area (as measured at the growth climate) was slightly lower in plants with poor nutrient availability, photosynthetic rate per unit leaf nitrogen was higher. These data suggest a priority of leaf nitrogen usage in photosynthesis, with limiting amounts of leaf nitrogen (and possibly other nutrients) for subsequent growth processes. This argument is consistent with the higher concentrations of starch found in plants with poor nutrient availability.     

STEPWISE EVOLUTION OF RESISTANCE TO TOXIC CARDENOLIDES VIA GENETIC SUBSTITUTIONS IN THE NA+/K+‐ATPASE OF MILKWEED BUTTERFLIES (LEPIDOPTERA: DANAINI)

Despite the monarch butterfly (Danaus plexippus) being famous for its adaptations to the defensive traits of its milkweed host plants, little is known about the macroevolution of these traits. Unlike most other animal species, monarchs are largely insensitive to cardenolides, because their target site, the sodium pump (Na⁺/K⁺‐ATPase), has evolved amino acid substitutions that reduce cardenolide binding (so‐called target site insensitivity, TSI). Because many, but not all, species of milkweed butterflies (Danaini) are associated with cardenolide‐containing host plants, we analyzed 16 species, representing all phylogenetic lineages of milkweed butterflies, for the occurrence of TSI by sequence analyses of the Na⁺/K⁺‐ATPase gene and by enzymatic assays with extracted Na⁺/K⁺‐ATPase. Here we report that sensitivity to cardenolides was reduced in a stepwise manner during the macroevolution of milkweed butterflies. Strikingly, not all Danaini typically consuming cardenolides showed TSI, but rather TSI was more strongly associated with sequestration of toxic cardenolides. Thus, the interplay between bottom‐up selection by plant compounds and top‐down selection by natural enemies can explain the evolutionary sequence of adaptations to these toxins.     

Photosynthesis and growth reduction with warming are driven by nonstomatal limitations in a Mediterranean semi‐arid shrub

Whereas warming enhances plant nutrient status and photosynthesis in most terrestrial ecosystems, dryland vegetation is vulnerable to the likely increases in evapotranspiration and reductions in soil moisture caused by elevated temperatures. Any warming‐induced declines in plant primary production and cover in drylands would increase erosion, land degradation, and desertification. We conducted a four‐year manipulative experiment in a semi‐arid Mediterranean ecosystem to evaluate the impacts of a ~2°C warming on the photosynthesis, transpiration, leaf nutrient status, chlorophyll content, isotopic composition, biomass growth, and postsummer survival of the native shrub Helianthemum squamatum. We predicted that warmed plants would show reduced photosynthetic activity and growth, primarily due to the greater stomatal limitation imposed by faster and more severe soil drying under warming. On average, warming reduced net photosynthetic rates by 36% across the study period. Despite this strong response, warming did not affect stomatal conductance and transpiration. The reduction of peak photosynthetic rates with warming was more pronounced in a drought year than in years with near‐average rainfall (75% and 25–40% reductions relative to controls, respectively), with no indications of photosynthetic acclimation to warming through time. Warmed plants had lower leaf N and P contents, δ¹³C, and sparser and smaller leaves than control plants. Warming reduced shoot dry mass production by 31%. However, warmed plants were able to cope with large reductions in net photosynthesis, leaf area, and shoot biomass production without changes in postsummer survival rates. Our findings highlight the key role of nonstomatal factors (biochemical and/or nutritional) in reducing net carbon assimilation rates and growth under warming, which has important implications for projections of plant carbon balance under the warmer and drier climatic scenario predicted for drylands worldwide. Projected climate warming over the coming decades could reduce net primary production by about one‐third in semi‐arid gypsum shrublands dominated by H. squamatum.     

Influence of vesicular arbuscular mycorrhizae and leaf age on net gas exchange of citrus leaves

The purpose of this study was to test the hypothesis that vesicular arbuscular mycorrhizal (VAM) fungi affect net assimilation of CO₂ (A) of different-aged citrus leaves independent of mineral nutrition effects of mycorrhizae. Citrus aurantium L., sour orange plants were grown for 6 months in a sandy soil low in phosphorus that was either infested with the VAM fungus, Glomus intraradices Schenck & Smith, or fertilized with additional phosphorus and left nonmycorrhizal (NM). Net CO₂ assimilation, stomatal conductance, water use efficiency, and mineral nutrient status for expanding, recently expanded, and mature leaves were evaluated as well as plant size and relative growth rate of leaves. Nutrient status and net gas exchange varied with leaf age. G. intraradices-inoculated plants had well-established colonization (79% of root length) and were comparable in relative growth rate and size at final harvest with NM plants. Leaf mineral concentrations were generally the same for VAM and NM plants except for nitrogen. Although leaf nitrogen was apparently sufficient for high rates of A, VAM plants did have higher nitrogen concentrations than NM at the time of gas exchange measurements. G. intraradices had no effect on A, stomatal conductance, or water use efficiency, irrespective of leaf age. These results show that well-established VAM colonization does not affect net gas exchange of citrus plants that are comparable in size, growth rate, and nutritional status with NM plants.     

CO2 enrichment reduces the relative contribution of latex and latex-related hydrocarbons to biomass in Euphorbia lathyris

The hypothesis that plants grown under elevated CO₂ allocate more carbon to the production of latex and C‐rich secondary compounds whereas nutrient addition counteracts this effect was tested. Two similar experiments were conducted in two different experimental facilities. In both facilities seedlings of Euphorbia lathyris were exposed to factorial combinations of two CO₂ concentrations and two levels of nutrient availability for 2 months. The CO₂ treatments and growth conditions differed substantially between these two experiments but treatment responses to elevated CO₂ and fertilizer addition were remarkably similar, underlining the robustness of our findings. Elevated CO₂ increased biomass to a greater extent in fertilized than in unfertilized plants and reduced the leaf biomass fraction by accelerating leaf senescence. Concentrations of non‐structural carbohydrates (NSC) increased in elevated CO₂. However, this apparent carbon surplus did not feed into the whole plant latex pool. The latex harvest per leaf (?25%) and the concentration of latex‐related hydrocarbons (?20%) even decreased under elevated CO₂ (both experiments P < 0.05). Fertilization reduced NSC concentrations (?25%) but neither affected latex yield per leaf nor the concentration of latex‐related hydrocarbons. It is concluded that latex and related hydrocarbons in CO₂‐enriched plants are a negligible sink for excess carbon irrespective of nutrient status and thus, vigour of growth.     

Phytochrome effects on leaf growth and chlorophyll content in Petunia axilaris

Abstract Changes in the phytochrome status at the end of the daily photosynthetic period result in several plant responses. To understand the causal relations among these responses it is useful to investigate species or experimental conditions where the most common correlations among responses are broken. A step in this direction is presented here with Petunia axilaris, where FR-treated plants showed low chlorophyll content and erect leaves, but- contrary to other species-higher leaf area and plant dry weight. Differences in area expansion were related to the late phase of leaf growth and were due, at least in part, to larger cells in FR-treated plants. Effects on length/width ratio, specific leaf area, net assimilation rate, shoot/root ratio and leaf number were small or non-existent. It is suggested that the lower chlorophyll content in FR-treated plants was not a consequence of scarcity of assimilates.     

Salinity and light interactively affect neotropical mangrove seedlings at the leaf and whole plant levels

We have studied the interactive effects of salinity and light on Avicennia germinans mangrove seedlings in greenhouse and field experiments. We hypothesized that net photosynthesis, growth, and survivorship rates should increase more with an increase in light availability for plants growing at low salinity than for those growing at high salinity. This hypothesis was supported by our results for net photosynthesis and growth. Net daily photosynthesis did increase more with increasing light for low-salinity plants than for high-salinity plants. Stomatal conductance, leaf-level transpiration, and internal CO₂ concentrations were lower at high than at low salinity. At high light, the ratio of leaf respiration to assimilation was 2.5 times greater at high than at low salinity. Stomatal limitations and increased respiratory costs may explain why, at high salinity, seedlings did not respond to increased light availability with increased net photosynthesis. Seedling mass and growth rates increased more with increasing light availability at low than at high salinity. Ratios of root mass to leaf mass were higher at high salinity, suggesting that either water or nutrient limitations may have limited seedling growth at high salinity in response to increasing light. The interactive effects of salinity and light on seedling size and growth rates observed in the greenhouse were robust in the field, despite the presence of other factors in the field—such as inundation, nutrient gradients, and herbivory. In the field, seedling survivorship was higher at low than at high salinity and increased with light availability. Interestingly, the positive effect of light on seedling survivorship was stronger at high salinity, indicating that growth and survivorship rates are decoupled. In general, this study demonstrates that environmental effects at the leaf-level also influence whole plant growth in mangroves.     

Seasonal cardenolide production and Dop5betar gene expression in natural populations of Digitalis obscura

Productivity variations and seasonal fluctuations of cardenolides have been studied in 10 natural populations of Digitalis obscura distributed in three bioclimatic belts. Main cardenolides in D. obscura plants are those of the series A and such predominance (ca. 80-85%) over the series B metabolites is independent of the population studied or the degree of maturity of the leaves. Primary glycosides represent ca. 50-60% of total cardenolides; this percentage did not vary among populations or with the leaf age but increased in summer and decreased in winter. A correlation analysis between plant biomass and cardenolide content showed a positive relationship of these parameters, which, according to the bioclimatic distribution of the populations, suggests that certain environmental conditions may cause marked decreases in plant biomass together with a reduction in productivity. Cardenolide contents changed in the timecourse of the four seasons as a multiple response to distinct plant and/or environmental factors. The lowest production was recorded in May, followed by a fast cardenolide accumulation in summer, a decreasing phase in autumn, and a stationary phase in winter. We also analysed the seasonal expression of the gene encoding the progesterone 5beta-reductase, enzyme producing the required 5beta-configured intermediaries of cardenolides. A fragment of the isolated partial genomic sequence was used as a probe for Northern analysis to study the seasonal gene expression in selected populations. The expression pattern showed increasing levels from February to July and a further reduction in autumn, although harmful climatic conditions seems to induce overexpression of this gene.     

Mammal Browsers and Rainfall Affect Acacia Leaf Nutrient Content,Defense, and Growth in South African Savannas

Five sets of herbivore exclosures situated in mesic and semi‐arid savannas in Hluhluwe‐iMfolozi Park, South Africa were used to investigate the effects of mammal browsers and savanna type on plant traits relating to leaf nutrient content, defense, and growth in seven Acacia species. Mostly, browsing did not significantly affect leaf nutrient content but for a few species (i.e., increasing foliar N and P, decreasing C/N, and total polyphenols). Browser effects on structural defenses tended to be more pronounced than for leaf nutrient content and chemical defenses, particularly for semi‐arid species, resulting in longer, thicker, and denser spines, and a lower bite size index on browsed plants for most semi‐arid species. Browsing had no significant effect on growth rates for all species. Secondly, we investigated the effect of savanna type (mesic vs. semi‐arid) on the same set of plant traits and growth rates. A trade‐off in defense strategy was evident where mesic species had lower quality leaves and invested more heavily in growth and chemical defenses, while semi‐arid species generally had higher nutrient content leaves and invested more in structural defenses and higher levels of ramification. These findings suggest that the previously documented trade‐off in plant growth, resprouting ability and architecture between herbivore versus fire‐adapted savanna woody species can possibly be extended to include browse quality and defense type.